DataSheet_2_High-Throughput Sequencing to Detect Novel Likely Gene-Disrupting Variants in Pathogenesis of Sporadic Brain Arteriovenous Malformations.pdf

Molecular signaling that leads to brain arteriovenous malformation (bAVM) is to date elusive and this is firstly due to the low frequency of familial cases. Conversely, sporadic bAVM is the most diffuse condition and represents the main source to characterize the genetic basis of the disease. Several studies were conducted in order to detect both germ-line and somatic mutations linked to bAVM development and, in this context, next generation sequencing technologies offer a pivotal resource for the amount of outputted information. We performed whole exome sequencing on a young boy affected by sporadic bAVM. Paired-end sequencing was conducted on an Illumina platform and filtered variants were validated by Sanger sequencing. We detected 20 likely gene-disrupting variants affecting as many loci. Of these variants, 11 are inherited novel variants and one is a de novo nonsense variant, affecting STK4 gene. Moreover, we also considered rare known variants affecting loci involved in vascular differentiation. In order to explain their possible involvement in bAVM pathogenesis, we analyzed molecular networks at Cytoscape platform. In this study we focus on some genetic point variations detected in a child affected by bAVM. Therefore, we suggest these novel affected loci as prioritized for further investigation on pathogenesis of bAVM lesions.

迄今，引发脑动静脉畸形（brain arteriovenous malformation，bAVM）的分子信号通路仍未明确，首要原因在于家族性病例的发生率极低。与之相对，散发性bAVM是最为普遍的病例亚型，亦是阐明该疾病遗传基础的核心研究对象。多项研究旨在检测与bAVM发生相关的生殖系及体细胞突变，在此背景下，下一代测序技术（next generation sequencing）可为海量产出数据的分析提供关键支撑。我们对1名罹患散发性bAVM的幼童开展了全外显子组测序（whole exome sequencing），采用Illumina平台完成双端测序（paired-end sequencing），并通过桑格测序（Sanger sequencing）对筛选后的变异进行验证。我们共检测到20个可能造成基因功能破坏的变异，分别累及20个基因座。其中11个为遗传性新发变异，另有1个为累及STK4基因的新生无义突变（de novo）。此外，我们还分析了累及血管分化相关基因座的罕见已知变异。为阐释这些变异在bAVM发病机制中的潜在作用，我们借助Cytoscape平台对分子网络进行了分析。本研究聚焦于1名bAVM患儿体内检测到的若干基因点变异。因此，我们建议将本次发现的受累基因座列为bAVM病灶发病机制后续研究的优先方向。